I am using primary VSMCs isolated from Abdominal aorta of knockout and transgenic mice. After passage 10, I noticed some changes in prolifration rate and expression of some protein.
What is the the recomended maximum passage for this kind of cells?
Using pulmonary artery smooth muscle cells (PASMC) from rat, that could be compared in some extent to the cells you are using, we used them around passage 3 or 4. Beyond this passage the cells started giving strange results, and even the shape was changing also. Maybe it could be due to changes in the phenotype. Anyhow, I wouldn´t recommend to use the cells at so high passages, if possible. To avoid having to prepare new primary cultures you could freeze alliquots when you are in an earlier passage, and defrost them when needed,
Good luck.
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